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Biomedical subjects

G J Gleich

Publications and source records attributed to G J Gleich.

At least 415 records · Page 23Linked to original sources
PubMed1977

Measurement of IgD by a double antibody radioimmunoassay: demonstration of an apparent trimodal distribution of IgD levels in normal human sera.

A double antibody radioimmunoassay (RIA) was developed to measure IgD in serum and secretions. One IgD myeloma protein was used as radiolabeled antigen and standard with antiserum to a second IgD myeloma protein. The IgD standard and normal sera yielded parallel inhibition curves in the RIA and inhibition was produced by IgD and not by any of the other immunoglobulins. The assay had a lower limit of sensitivity of 0.01 International Unit (I.U.)/ml and modifications increased the sensitivity to 0.0008 I.U./ml. Measurable IgD levels were found in all 112 normal adult sera assayed (geometric mean 13.0 I.U./ml, arithmetic mean 30.1 I.U./ml, median 14.8 I.U./ml, range 0.10 to 202 I.U./ml, 95% of values between 0.19 and 156 I.U./ml. The distribution of IgD in the 112 normal sera appeared trimodal with modes at approximately 0.25 I.U./ml, 5I.U./ml, and 35 I.U./ml. IgD was measurable in nasal and bronchial washes and human milk, but could not be detected in parotid fluid.

Antigens↗
PubMed1976

Atopic dermatitis and IgE. Relationship between changes in IgE levels and severity of disease.

At the Mayo Clinic, atopic dermatitis was evaluated in a series of 28 patients to determine whether, during the course of the disease, serum IgE levels fluctuated with changes in the severity of dermatitis. The study covered a 1- to 11/2-year follow-up period. Previously, it had been shown that, at initial examination, serum IgE levels were positively related to the extent of disease. Although notable fluctuations in serum IgE levels and in extent of dermatitis were seen in more than half of the cases, no relationship was found between the two factors during the period of study.

Dermatitis, Atopic↗
PubMed1976

Studies on Alternaria allergens. I. Establishment of the radioallergosorbent test for measurement of Alternaria allergens.

The ability to covalently couple Alternaria allergens to microcrystalline cellulose particles has permitted not only the measurement of IgE antibodies to Alternaria in patient serums but also the identification of allergenic fractions from crude Alternaria extracts. Crude aqueous Alternaria extracts from 3 commerical suppliers were coupled to cellulose but failed to bind more than 5% of total radioactive counts (TRC) when reacted with serums from highly sensitive patients. Fractionation of a commercial extract through Sephadex G-25 showed that almost all allergenic activity was located in a protein- and carbohydrate-containing peak eluting at the column void volume. These fractions were pooled and coupled to cellulose to yield a RAST polymer which produced up to 20% TRC binding when tested with serums from over 100 Alternaria-sensitive patients, and only up to 1% TRC binding with 17 nonallergic serums. The study of commercial Alternaria extracts by chromatographic and Rast inhibition techniques showed that present extracts are neither qualitatively or quantitatively comparable.

Allergens↗
PubMed1976

Detection of allergy to nuts by the radioallergosorbent test.

The diagnosis of food allergy is often difficult to make by conventional means. Histories are frequently ambiguous, and skin testing is of dubious reliability because of the number of false-positive and false-negative reactions. We have evaluated the radioallergosorbent test (RAST) for the in vitro measurement of the specific IgE antibodies to nuts, including Brazil nut, almond, walnut, pecan, cashew, and the legume, peanut. Serums were obtained from 18 patients with a history of nut allergy and IgE level and specific IgE antibodies were measured. Thirteen of the 18 patients had significantly elevated IgE antibody (greater than twice control) to one or more of the allergens. Prausnitz-Küstner tests on selected serums in general corroborated the results of the in vitro studies. Five patients had RAST elevations to 2 or more nuts. As a group RAST-positive patients had elevated mean serum IgE levels and more severe clinical symptoms (p less than 0.01). The specificity and cross-reactivity of IgE antibodies to different nut antigens was investigated by RAST inhibition with serums from 5 patients having high levels of IgE antibody. In 4 patients no cross-reactivity between Brazil nut and peanut was found. In contrast, several nut extracts inhibited the reaction of pecan allergen with IgE antibodies. These results indicate that specific IgE antibodies can be measured by RAST in patients with nut allergy and the cross-reactivity of nut antigens can be investigated. RAST would appear to be most useful in confirming the diagnosis of nut hypersensitivity in children or in highly allergic patients in whom skin testing poses a risk of anaphylaxis.

Absorption↗
PubMed1976

Analysis of the potency of extracts of June grass pollen by their inhibitory capacities in the radioallergosorbent test.

The potencies of 11 commercial extracts of June grass pollen were analyzed by skin test end point titrations and compared to potencies as determined in vitro (1) by the radioallergosorbent test (RAST), (2) by Group I antigen content, and (3) by protein nitrogen units (PNU). RAST potencies were determined by the capacity of the extract to inhibit the binding of IgE antibody to solid-phase allergen, and they were expressed as the quantity of extract required for 50% inhibition of binding. Potencies determined by skin testing in 8 patients were significantly related among the various patients in 19 of 27 comparisons and showed differences of up to 95,000-fold in the strengths of the extracts. Estimation of potencies by RAST inhibition showed approximately a 100-fold difference among the extracts and in 5 of 8 cases these were significantly related to potencies measured by skin tests. Similarly, PNU determinations and Group I determinations were also significantly related to potencies by skin test titration in 5 of 8 and in 4 of 8 comparisons, respectively. Comparison of the geometric mean skin test potencies with RAST, PNU, and Group I potencies revealed that all were significantly related to skin test potencies although the correlation of RAST and skin potency was the highest. The results indicate that measurement of potency by RAST inhibition compares favorably with other in vitro measurements of potency. These results are compared with those of a prior study with extracts of short ragweed, and the reasons for the differences between the results in the two studies are discussed.

Absorption↗
PubMed1976

Studies on Alternaria allergens. II. Measurement of the relative potency of commercial Alternaria extracts by the direct RAST and by RAST inhibition.

The relative potency of 12 commercial Alternaria extracts was analyzed by end-point skin test titrations and compared to in vitro measurements of potency, including (1) the radioallergosorbent test (RAST) inhibition procedure; (2) the direct RAST procedure; and (3) the protein nitrogen unit (PNU) content. Potencies determined by skin testing 10 sensitive patients were strongly correlated among the various patients. Measurements of potency by both RAST inhibition and direct RAST assay were strongly correlated to potency as measured by skin testing. In contrast, neither the weight:volume nor the PNU content bore any relationship to allergenic potency as measured by skin testing or by either of the RAST procedures. Extracts differed by as much as 3,000-fold in allergen content by skin testing. Moreover, the extracts appeared to contain different allergenic determinants when tested by RAST inhibition. RAST inhibition offered several technical advantages over the direct RAST procedure, in that only one solid-phase RAST reagent was required, slopes of dose-response curves could be more easily compared, and a greater discrimination in allergenic potencies among extracts could be made. The RAST appears to offer an excellent method for measuring the potency of allergy extracts, pending the isolation and characterization of actual allergens.

Allergens↗
PubMed1976

The cross-reactivity of IgE antibodies with pollen allergens. I. Analyses of various species of grass pollens.

Atopic patients with histories of grass pollen allergy often are sensitive to a variety of species of grasses. Using a serum pool from patients sensitive to June grass, we analyzed the reactivity of IgE antibodies to seven grasses by the radioallergosorbent test. Extracts were analyzed for their inhibitory activities with solid-phase allergens prepared from all of the grass pollen. Also samples of serum were exhaustively absorved with solid-phase allergens and the supernatants tested to determine the reactivity of the remaining IgE antibodies. Three patterns of reactivity were observed: (1) June, orchard, meadow fescue, and perennial rye grasses displayed similar reactivity in both inhibition and absorption studies; (2) sweet vernal and Bermuda grasses were considerably less reactive with the serum pool, indicating that they lacked antigenic determinants possessed by the other grasses; and (3) timothy grass possessed unique antigenic determinants. Knowledge of these patterns of cross-allergenicity is of importance for diagnosis and treatment of sensitive patients as well as for in vitro standardization of extracts.

Absorption↗
PubMed1976

The cross-reactivity of IgE antibodies with pollen allergens. II. Analyses of various species of ragweed and other fall weed pollens.

Although the antigenic composition of short ragweed pollen has been extensively investigated, very few studies have analyzed the allergenic cross-reactivity among various types of ragweed pollens. Using a serum pool from patients sensitive to short ragweed, we studied the cross-reactivity of IgE antibodies to six ragweeds by the radioallergosorbent test. Extracts were analyzed for their inhibitory activities with solid-phase allergens prepared from all of the ragweed pollens. Also, samples of serum were absorbed with the various solid-phase allergens and the reactivity of the remaining IgE antibodies was determined. Two patterns of reactivity were observed. Short, giant, western, and false ragweeds displayed comparable reactivity in both inhibition and absorption experiments. Slender and southern ragweed were considerably less active, indicating that they lacked allergenic groupings possessed by the other species. These same patterns of cross-reactivity were found using ragweed pollens from four commercial sources. Knowledge of these patterns of cross-allergenicity is of importance for diagnosis and treatment of sensitive patients as well as for in vitro standardization of extracts.

Absorption↗
PubMed1976

Raised serum immunoglobulin E in Wegener's granulomatosis.

Five patients with Wegner's granulomatosis were found to have significantly raised serum immunoglobulin E (IgE) levels. The rise in IgE was not related to the extent of clinical involvement, was not part of a generalized serum immunoglobulin rise, and was not associated with eosinophilia. Raised serum IgE may be a clue to the pathogenesis of this disease.

Adult↗
PubMed1976

Comparative properties of the Charcot-Leyden crystal protein and the major basic protein from human eosinophils.

Guinea pig eosinophil granules contain a protein, the major basic protein (MBP), which accounts for more than half of the total granule protein, has a high content of arginine, and displays a remarkable tendency to form disulfide-linked aggregates. In this study we have purified a similar protein from human eosinophil granules and have compared the human MBP to the protein comprising the Charcot-Leyden crystal (CLC). Eosinophils from patients with various diseases were purified and disrupted, and the granule fraction was obtained. Examination of the granule fraction by transmission electron microscopy showed numerous typical eosinophil granules. Analyses of granule lysates by gel filtration and by polyacrylamide gel electrophoresis revealed the presence of peroxidase and MBP with properties similar to that previously found in guinea pig eosinophil granules. The human MBP had a molecular weight of 9,200, contained less than 1% carbohydrate, was rich in arginine, and readily formed disulfide-bonded aggregates. CLC were prepared from eosinophil-rich cell suspensions by homogenization in hypotonic saline. The supernates following centrifugation of cell debris spontaneously formed CLC. Analysis of CLC revealed the presence of a protein with a molecular weight of 13,000 containing 1.2% carbohydrate. The protein displayed a remarkable tendency to aggregate even in the presence of 0.2 M acetic acid. Human MBP and CLC protein differed in their molecular weights, carbohydrate compositions, and amino acid analyses. Mixtures of the MBP and the CLC protein yielded two bands in polyacrylamide gel electrophoresis. Neither eosinophil protein increased vascular permeability in the guinea pig skin or contracted the guinea pig ileum. The results indicate that the human MBP and the CLC are distinct substances with properties such that one cannot be derived from the other.

Animals↗
PubMed1976

The late phase of the immediate wheal and flare skin reaction. Its dependence upon IgE antibodies.

IgE antibodies are usually thought to induce only immediate skin reactions. We have shown that the intradermal injection of a number of different allergens can produce a prolonged inflammatory reaction after the immediate wheal and flare in most sensitive subjects. This late inflammatory response occurs 6-12 h after challenge and is characterized by diffuse edema, erythema, pruritus, and heat. Both immediate and late responses can also be seen after passive sensitization of skin sites in nonatopic subjects. That IgE is involved in inducing the reaction was shown by the abolition of both immediate and late responses by passive transfer tests in the following experiments: (a) heating atopic serum at 56degreesC for 4 h, (b) removing IgE from the atopic serum by a solid phase anti-IgE immunoabsorbent, and (c) competitively inhibiting the binding of IgE antibodies to cells by an IgE myeloma protein. In addition, both responses were induced by affinity chromatography-purified IgE antibody, followed by antigenic challenge. Very similar lesions could also be induced by intradermal injection of Compound 48/80, thus suggesting a central role in the reaction for the mast cell or basophil. Histologically, the late phase is characterized by edema and a mixed cellular infiltration, predominantly lymphocytic but also containing eosinophils, neutrophils and basophils. Direct immunofluorescent staining did not show deposition of immunoglobulins or complement components, except IgM in 2 of 15 and C3 in 1 of 15 patients. This finding indicates that the late phase does not depend on the deposition of immune complexes. The results of the study suggest that IgE-allergen interaction on the surfaces of mast cells or on infiltrating basophils causes both immediate and late cutaneous responses.

Adult↗
PubMed1976

Isolation and partial characterization of a major basic protein from rat eosinophil granules.

The rat eosinophil granule possesses a major basic protein (MBP) similar to that previously isolated from human and guinea pig eosinophils. This conclusion is based on demonstration that the major protein of the rat eosinophil granule is a low molecular weight, highly basic material with an amino acid composition similar to that of the human and guinea pig MBP.

Amino Acids↗
PubMed1976

Endomyocardiopathy with eosinophilia.

Five patients were seen at the Mayo Clinic over an 8-year period with the following complex of clinical and morphologic features; striking eosinophilia, cardiomyopathy, hepatosplenomegaly, and either a rapidly fatal or a prolonged, debilitating illness. In recent years, controversy has raged over the precise designation of this syndrome, with proposals ranging from eosinophilic leukemia to hypereosinophilic syndromes. To focus on the major target organ of the disease, we have favored the term endomyocardiopathy with eosinophilia. Experience with these five patients showed that (1) eosinophilia can persist for many years before symptoms appear; (2) progressive restrictive cardiac disease was the major cause of death and debility; (3) osmiophilic cytoplasmic inclusions are present in eosinophils of these patients and also in cells from other patients with marked eosinophilia; and (4) echocardiography may prove to be a useful noninvasive tool to diagnose and follow the progress of cardiac involvement. Although none of these patients was thought to have leukemia, intensive therapy with steroids or cytotoxic agents, or both, is considered necessary to control the progression of the disease.

Adolescent↗
PubMed1975

Immunoglobulin E antibodies to pollen allergens account for high percentages of total immunoglobulin E protein.

The quantities of immunoglobulin E (IgE) antibodies to grass or ragweed allergens were measured by an immunoabsorption in the serums of patients sensitive to one of these allergens. IgE antibodies to grass or ragweed allergens accounted for means of 30 and 29 percent of the total IgE protein. After the ragweed pollination season, the levels of serum IgE antibodies to ragweed allergens rose dramatically and in postpollination serums they accounted for 39 percent of the total IgE protein with a range from 13 to 50 percent.

Humans↗
PubMed1975

Measurement of IgE antibodies by the radioallergosorbent test. I. Technical considerations in the performance of the test.

Since its introduction by Wide, Bennich, and Johansson in 1967, the radioallergosorbent test (RAST) has been increasingly used for measurement of specific IgE antibodies. In this study we have varied the conditions during the performance of the RAST and have investigated the effect of altering: (1) the speed of rotation of tubes, (2) the number of washes in both steps of the test, (3) the length of rotation of tubes, (4) the temperature, (5) the conditions during centrifugation, (6) the quantity of antibody to IgE in the second step of the test, (7) the composition of buffers in the reactions, (8) the type of tube employed in the test, (9) the quantity of antigen employed during the coupling procedure, and, finally, (10) the iodine isotope, 125I or 131I, used to radiolabel anti-IgE. Of these variables the most significant were the quantity of anti-IgE used in the second step of the test and the quantity of antigen used in the coupling procedure. Other variables of importance were the choice of radioiodine, whether the tubes were rotated or not, the type of tube used, and finally the diluent employed.

Allergens↗
PubMed1975

Measurement of IgE antibodies by the radioallergosorbent test. II. Analyses of quantitative relationships in the test.

IgE antibodies have been measured by the radioallergosorbent test (RAST) and the relative amounts present in serum determined by comparison with reference standards. In this study we analyzed the quantitative aspects of the binding of IgE antibody to solid-phase ragweed antigens. With the volumes of allergic serum usually tested, IgE antibodies are in excess in the first step of the RAST. The inability of solid-phase antigens to remove IgE antibody appeared to be due to an insufficient quantity of antigen on the particles rather than steric interference with binding of IgE antibody. Dose-response curves with serums from several ragweed-sensitive subjects were not parallel when plotted on a semilog scale. In contrast, log-log plots of dose-response curves with serums from ragweed-sensitive subjects were nearly parallel. Log-log plots of dose-response curves with serums from subjects with ragweed and grass sensitivity and tested with the appropriate solid-phase antigens also were nearly parallel. Because of the latter finding, RAST could be standardized using a reference serum in every assay and plotting the results on a log-log scale. Finally, because IgE antibody is in excess in the RAST as it is usually performed, the final result reflects both the quantity and the affinity of the IgE antibody.

Allergens↗
PubMed1975

The effect of cromolyn sodium powder as a treatment for ragweed pollinosis.

The effect of treating ragweed pollinosis with intranasal cromolyn sodium powder was evaluated in 26 patients in a double-blind clinical study. Thirteen matched patient pairs were treated by either nasal insufflation of cromolyn sodium powder, 20 mg three times daily, or by a placebo powder. Patients were monitored for severity of symptoms by means of daily symptom diaries collected weekly throughout the pollen season. Serum samples were obtained in July prior to the ragweed season and following the ragweed season during the first and fourth weeks of October for the determination of ragweed-specific IgE antibody levels by the radioallergosorbent test. A significant reduction in symptom severity was observed in the treated patients with high preseasonal levels of IgE antibody to ragweed. Initially the levels of IgE antibody to ragweed did not differ between the two groups. IgE antibody levels increased in both groups after pollen exposure, but the treated group showed a significantly greater rise in IgE antibody levels. The results suggest that treatment with cromolyn sodium enhanced IgE antibody response to pollen exposure and significantly reduced the severity of hay fever symptoms in patients with high preseasonal levels of IgE antibody to ragweed.

Administration, Intranasal↗